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Transient MicroRNA Expression Enhances Myogenic Potential of Mouse Embryonic Stem Cells

机译:瞬时microRNA表达增强了小鼠胚胎干细胞的肌原源性潜力

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摘要

MicroRNAs (miRNAs) are known regulators of various cellular processes, including pluripotency and differentiation of embryonic stem cells (ESCs). We analyzed differentiation of two ESC linesD3 and B8, and observed significant differences in the expression of miRNAs and genes involved in pluripotency and differentiation. We also examined if transient miRNA overexpression could serve as a sufficient impulse modulating differentiation of mouse ESCs. ESCs were transfected with miRNA Mimics and differentiated in embryoid bodies and embryoid body outgrowths. miRNAs involved in differentiation of mesodermal lineages, such as miR145 and miR181, as well as miRNAs regulating myogenesis (MyomiRs)miR1, miR133a, miR133b, and miR206 were tested. Using such approach, we proved that transient overexpression of molecules selected by us modulated differentiation of mouse ESCs. Increase in miR145 levels upregulated Pax3, Pax7, Myod1, Myog, and MyHC2, while miR181 triggered the expression of such crucial myogenic factors as Myf5 and MyHC2. As a result, the ability of ESCs to initiate myogenic differentiation and form myotubes was enhanced. Premature expression of MyomiRs had, however, an adverse effect on myogenic differentiation of ESCs.
机译:MicroRNAS(miRNA)是各种细胞过程的已知调节因子,包括胚胎干细胞的多能和分化(ESC)。我们分析了两个ESC线3和B8的分化,并观察到麦芽糖和参与多能性和分化的基因表达的显着差异。我们还检查了瞬态miRNA过表达是否可以用作小鼠ESC的足够脉冲调制分化。将ESC用miRNA模拟转染并在胚胎体和胚胎体外分化。研究了MiR145和MiR181等介质谱系的分化的miRNA,以及调节肌瘤的miRNA(myomirs)miR1,miR133a,mir133b和mir206。使用这种方法,我们证明了由美国鼠标ESC的调节分化选择的分子的瞬时过度表达。增加miR145级别上调PAX3,PAX7,MyoD1,MYOG和MYHC2,而MIR181则引发如MYF5和MYHC2的这种关键的源性因子的表达。结果,增强了ESC以引发肌遗传分化和形成肌管的能力。然而,Myomirs的过早表达对ESC的肌遗传分化产生不利影响。

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